Expression, purification, crystallization and preliminary crystallographic study of the SRA domain of the human UHRF1 protein.
Human UHRF1 belongs to the unique mammalian family of proteins which contain a SET- and RING finger-associated (SRA) domain. This 180-residue domain has been reported to play key roles in the functions of the protein. It allows UHRF1 to bind methylated DNA, histone deacetylase 1 and DNA methyltransferase 1, suggesting a bridge between DNA methylation and the histone code. No structural data is available for any SRA domain. Native and SeMet-labelled SRA domains of human UHRF1 were overexpressed in Escherichia coli cells, purified to homogeneity and crystallized using the hanging-drop vapour-diffusion method. A complete MAD data set was collected to 2.2 A resolution at 100 K. Crystals of the SeMet-labelled protein belonged to the trigonal space group P3(2)21, with unit-cell parameters a = b = 53.78, c = 162.05 A.
Delagoutte
Bénédicte
B
Département de Biologie et de Génomique Structurales, IGBMC, CNRS/INSERM Université Louis Pasteur, Parc d'Innovation BP 10142, 67404 Illkirch CEDEX, France.
Lallous
Nada
N
Birck
Catherine
C
Oudet
Pierre
P
Samama
Jean Pierre
JP
D022762:CCAAT-Enhancer-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D003460:Crystallization; D003461:Crystallography / Q000379:methods; D004247:DNA / Q000378:metabolism; D019175:DNA Methylation; D006655:Histone Deacetylases / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D006801:Humans; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D011485:Protein Binding; D044767:Ubiquitin-Protein Ligases
D022762:CCAAT-Enhancer-Binding Proteins; D006657:Histones; D010446:Peptide Fragments; D004247:DNA; C406364:UHRF1 protein, human; D044767:Ubiquitin-Protein Ligases; D006655:Histone Deacetylases